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Egg quality in furnished cages for laying hens--effects of crack reduction measures and hybrid.

Egg quality, egg production, and hen use of facilities, with respect mainly to proportion of eggs laid in nests and bird locations after lights-out, were studied in furnished-cage models for six or eight birds. In these cages, most of the eggs are laid in the 25 cm wide nest, i.e., egg laying is concentrated to a much smaller area compared with conventional cages. The study (20 to 80 wk of age) used 1,296 hens of three genotypes Hy-Line White, Hy-Line Brown, and Lohmann Selected Leghorn. Conventional four-hen cages were included as a reference. We separately studied the effects on the proportion of eggs cracked by egg saver wires in front of the egg cradle, long nest curtains, and position of nest opening (rear or front) with 30 or 100% of the nest bottom being lined with AstroTurf. Egg saver wires and long nest curtains lowered the proportions of cracked eggs significantly by softly catching and reducing the speed of the eggs before entering the egg cradle. The position of the nest opening affected none of the measured parameters. Less lining in the bottom of the nest, i.e., a reduction to 30% coverage, resulted in significantly higher proportions of dirty eggs and lower proportions of eggs laid in nests. Hybrid differences were found in most of the measured traits. We concluded that devices like egg savers and long nest curtains are effective measures in reducing cracks in furnished cages, where egg laying is concentrated to a much smaller area than in conventional cages. Reduction of the bottom lining to 30% makes nests less attractive to birds laying eggs.

Animals↗

Mapping of quantitative trait loci affecting quality and production traits in egg layers.

A mapping population segregating for egg quality traits was created by a line cross between two egg layer lines and screened by a genome scan. The F2 generation consisted of 307 hens, which were scored for egg quality and production traits. The mapping population was genotyped for 99 microsatellite loci, spanning nine macrochromosomes and five small linkage groups. The linkage maps were used in mapping QTL affecting 14 traits, by using multiple markers and a least-squares approach. We detected 14 genomewide significant and six suggestive QTL that were located on chromosomes 2, 3, 4, 5, and, 8 and sex chromosome Z. A significant QTL affecting egg white thinning was found on chromosome 2. For eggshell strength, a significant QTL was found on chromosome Z. For production traits, the most interesting area was on chromosome 4, where highly significant QTL effects were detected for BW, egg weight, and feed intake in the same area. The most significant QTL explains 25.8% of the phenotypic variance in F2 of body weight. An area affecting the age at first egg, egg weight, and the number of eggs was located on chromosome Z.

Animals↗

The use of densitometry to detect differences in bone mineral density and content of live White Leghorns fed varying levels of dietary calcium.

Densitometry was investigated as a noninvasive tool to monitor skeletal integrity in live White Leghorns as an indicator for osteoporosis, a noninfectious disease resulting in mineral loss from the bone. The objectives of the experiment were 1) to assess the ability of densitometry to detect differences in bone integrity in live White Leghorns fed varying concentrations of dietary calcium and 2) to correlate densitometric scans with other bone test methods and production parameters that are sensitive to calcium concentrations in the diet. Hens were fed hypercalcemic (5.4%), control (3.6%), or hypocalcemic (1.8%) diets from 32 to 58 wk of age. A Norland densitometer was used to assess bone mineral density (BMD) and bone mineral content (BMC) of the left tibia and humerus in restrained, unanesthetized hens at 36, 46, and 56 wk of age (experiment 1) and at 38, 48, and 58 wk of age (experiment 2). Bones were excised from hens at 38, 48, and 58 wk of age for breaking strength measurements. Results from the densitometric scans showed that BMD and BMC of the humerus and tibia of live hens decreased linearly when hens consumed diets with decreasing concentrations of calcium (experiment 2). Similar trends in BMD and BMC were detected in experiment 1 at 36 wk of age using BW as a covariate. The results from the densitometric scans were comparable to those obtained from other bone tests commonly used. For example, bone breaking force, stress, and modulus of elasticity decreased linearly as hens consumed decreasing concentrations of calcium. Bone breaking force was correlated with BMD (r=0.65, P<0.001). We concluded that densitometry accurately measures differences in BMD and BMC in live birds fed varying concentrations of dietary calcium.

Absorptiometry, Photon↗

Assessing bone mineral density in vivo: quantitative computed tomography.

Egg-laying hens require substantial amounts of Ca to support eggshell formation. Over time, structural bone is catabolized to provide some of the Ca required; the structural bone is not replaced. As the hen ages, this can eventually lead to osteoporosis. Quantitative Computed Tomography (QCT) is a nondestructive technique used to measure bone mineral density (BMD). QCT is used diagnostically in humans to assess osteoporosis; BMD determined by QCT is correlated with other, more invasive methods of bone mineral determinations, such as ashing. An x-ray is sent through a bone at multiple angles within a plane to generate a 2-dimensional image and a 3-dimensional calculation of volume and BMD. The technique allows resolution of total, trabecular, and cortical BMD and cross-sectional areas. The separation of bone types allows very precise measurements of the bone compartments most important in Ca supply for eggshell formation and bone strength. QCT has been adapted in our laboratory to measure BMD in vivo and ex vivo in poultry; values obtained for poultry bones are moderately correlated with destructive means of assessing bone quality such as breaking strength, ashing, and chemical bone mineral determinations. Thus, changes in BMD of individual birds can be measured over time; BMD at specific time points can be correlated with production parameters and eggshell quality traits. QCT is an effective technique to measure BMD in laying hens, which allows resolution of total BMD as well as cortical and trabecular BMD.

Aging↗

Effect of vitamin D2- and D3-enriched diets on egg vitamin D content, production, and bird condition during an entire production period.

Vitamin D insufficiency during winter is a common problem for humans in Europe. One way to ease this problem is through the production of vitamin D-fortified eggs. To evaluate such a production process, the effects of vitamin D supplementation during an entire production period were assessed. Transfer of vitamin D3 (cholecalciferol) and vitamin D2 (ergocalciferol) from the diet to egg yolks was measured using 2 different levels of both vitamins (6,000 and 15,000 IU/kg feed) relative to a control treatment (2,500 IU vitamin D3/kg feed). During the experiment, production parameters, egg quality (egg weight, Haugh unit, specific gravity, eggshell fracture force, and Ca content of eggshell), and the condition of hens were monitored. At the end of the experiment histopathological tests were performed. Supplementing diets with vitamin D3 increased egg yolk vitamin D content more effectively than did supplementation with vitamin D2. For groups of hens receiving 6,000 or 15,000 IU of vitamin D3/kg feed, egg yolk vitamin D3 content ranged from 9.1 to 13.6 and from 25.3 to 33.7 microg/100 g, respectively. Corresponding values for birds fed vitamin D2 were 4.7 to 7.0 and 13.3 to 21.0 microg/100 g. Both supplements enhanced vitamin D3 content of egg yolks relative to the control diet (2.5 to 5.0 microg/100 g of egg yolk). Vitamin D supplements had no effects on production parameters compared with the control diet. However, especially vitamin D3 improved bone strength (P < 0.05). Autopsy at the end of the experiment indicated no detrimental accumulation of calcium in the kidneys, liver, heart, muscles, or lungs.

Animals↗

Effects of S6-strain Mycoplasma gallisepticum inoculation at ten, twenty-two, or forty-five weeks of age on the performance characteristics of commercial egg laying hens.

Experimental inoculation of commercial laying hens, maintained under controlled conditions, with the S6-strain of Mycoplasma gallisepticum (S6MG) at 10 wk of age has previously been shown to affect the lengths and weights of various portions of the reproductive tract without affecting subsequent performance. Two trials were conducted to compare the effects of S6MG inoculation at 10 wk of age (prior to lay), 22 wk of age (onset of lay), and 45 wk of age (during lay) on performance characteristics in commercial layers housed and maintained under controlled conditions, as in previous studies. In each trial, BW, mortality, egg production, egg weight, eggshell weight per unit of surface area, percentage eggshell weight, percentage albumen weight, percentage yolk weight, and yolk weight per albumen weight ratio were examined at various ages throughout an entire laying cycle. Across wk 47 and 58 (age periods after the last 45 wk inoculation), eggshell weight per unit of surface area and percentage eggshell weight were significantly reduced in birds that had received an S6MG inoculation at 45 wk of age when compared with birds that had not received an S6MG inoculation or had been inoculated with S6MG at either 10 or 22 wk of age. Alterations in eggshell quality in response to S6MG may become evident only in older birds that are experiencing declines in production when housed under controlled conditions.

Aging↗

Effects of a dried Bacillus subtilis culture on egg quality.

The effects of a dried Bacillus subtilis culture on the egg qualities of layers were studied. Nine hundred and sixty 25-wk-old Lohmann Brown layers were randomly divided into 5 groups with 192 layers in each group. Layers in group 1 were fed a control diet. The remaining groups received the control diet that contained either 20 mg of zinc bacitracin/kg and 4 mg of colistin sulfate/kg or 500, 1,000, or 1,500 mg of B. subtilis culture/ kg, respectively. The results showed improvements in egg production, feed consumption, and feed conversion (P < 0.05) of layers when 500 mg of B. subtilis culture/kg was added to the diets. The results also showed some special improvements in this group, including increases in eggshell thickness, yolk color, and Haugh unit, and decreases in yolk cholesterol concentration (P < 0.05). However, excessive doses of B. subtilis culture did not improve the performance of layers.

Animals↗

Precopulatory mate guarding and mating behaviour in the rotifer Epiphanes senta (Monogononta: Rotifera).

Epiphanes senta is a littoral rotifer species that occurs in temporary waters and displays a mating behaviour which has not, to my knowledge, so far been described for monogonont rotifers. Monogonont rotifers show distinctive periods within their life cycle during which mictic females appear. Mictic females produce haploid eggs that develop into males or into diapausing eggs if fertilized. The females of E. senta are mostly stationary on the substrate while males are more active swimmers. If they encounter eggs with female embryos of their own species, they attend them and mate with the hatching female. Experiments showed that males are able to discriminate between male, female and diapausing eggs. They exhibit a strong preference for female eggs that are only a few hours away from hatching compared with eggs in early developmental stages. Further experiments did not show any significant differences in male attendance of mictic and amictic eggs. It is hypothesized that males judge the age of a female egg by sensing a chemical that is produced by the growing embryo and diffuses through the egg shell. The male mating behaviour is similar to precopulatory mate guarding known from arthropods but it lacks the monopolization of the female by the male.

Analysis of Variance↗

Early development and embryology of the platypus.

Information on the pre-hatching development of the platypus, Ornithorhynchus anatinus, is reliant on a small number of specimens, whose precise age is unknown. Material collected for J. P. Hill and now housed in the Hubrecht International Embryological Laboratory, Utrecht, contributes a major source of specimens. This paper presents new observations on developmental stages from the Hill collection, which allow for a more complete description of pre-hatching development. A feature of the pre-embryonic development of the platypus is the incomplete meroblastic cleavage. A column of fine yolk spheres extends from beneath the embryonic blastodisc towards the centre of a yolky vitellus, as seen in birds. The major expansion of extra-embryonic membranes occurs after the formation of the primitive streak. The primitive streak develops within an embryonal area as part of the superficial wall of the yolk-sac, a feature also shared with marsupials, birds and reptiles. The full-term, subspheroidal, intrauterine egg of the platypus has a major axis of about 17 mm and contains a flat, 19-20 somite, neurula-stage embryo which has prominent trigeminal ganglion primordia. The embryo at this stage is in a period of rapid modelling of the major early organ primordia of the nervous system, cardiovascular system, excretory system, and somite-derived components of the body wall. Soon after laying, five primary brain vesicles are present, the trigeminal ganglia CN5 as well as CN7, CN8, CN9, CN10, CN11 and CN12 are well developed. The alimentary system has an expanded stomach, pancreatic primordia and a gall bladder. Mesonephric tubules are associated with patent mesonephric ducts, which empty laterally into the cloaca. Extra-embryonic membranes at this stage show an extensive chorioamniotic connection that extends through the greater part of the caudal half of fused amniotic folds. The vascularized yolk-sac consists of a superficial yolk-sac omphalopleura and a deep yolk-sac splanchnopleure. The non-vascularized yolk-sac comprises one-quarter of the ahembryonal pole. Some distinctive monotreme features have developed by the mid-incubation period. The head is bent at an acute angle to the main body axis. The blunt upturned snout marks the site of the future oscaruncle and on the maxilla there is a median primordial papilla representing the egg tooth. The eye is open with a partly pigmented retinal ring. The forelimbs have partly separated digits, and the hindfeet are paddles. Just before hatching the upturned snout contains an oscaruncle and a sharp recurved median egg tooth. Forelimbs are pronated with separate digits possessing claw primordia. Portions of the highly vascularized extra-embryonic membranes are attached to the umbilical region and the flattened vesicular allantois has a distal region fused with the chorion. Prominent features of the hatchling are the presence of a bluntly conical oscaruncle and a translucent, horn-like egg tooth. These structures are though to enable the hatchling to extricate itself from the egg shell. At hatching, the forelimbs exhibit clawed digits and are capable of digitopalmar prehension. Hindlimbs are still paddles with digital rays. A prominent yolk-sac navel is present. The newly hatched platypus has an external form similar to that of a new-born marsupial. The early development of the platypus has many major differences to the developmental sequence for humans, which has been categorized by the use of Carnegie Stages. The rate of somitogenesis of the platypus is faster in relation to the central nervous system morphogenesis than seen in humans, and the size of the early platypus embryonal area is massive in relation to that of humans. The unique morphology and function of extra-embryonic membranes in the platypus defies comparative staging with human development. Structures adapted for altricial survival of the platypus hatchling require the acquisition of functional competence at an earlier stage of organogenesis than seen in eutherians, although they are reminiscent of those found in new-born marsupials.

Allantois↗

Flexibacter ovolyticus sp. nov., a pathogen of eggs and larvae of Atlantic halibut, Hippoglossus hippoglossus L.

A psychrotrophic Flexibacter sp., Flexibacter ovolyticus sp. nov., was isolated from the adherent bacterial epiflora of Atlantic halibut (Hippoglossus hippoglossus L.) eggs and was shown to be an opportunistic pathogen for halibut eggs and larvae. The strains which we isolated had the enzymatic capacity to dissolve both the chorion and the zona radiata of the egg shells. A total of 35 isolates were characterized by using morphological and biochemical tests. These strains were rod shaped, gram negative, Kovacs oxidase positive, and pale yellow and exhibited gliding motility. They did not produce acid from any of the wide range of carbohydrates tested. Our isolates had the ability to degrade gelatin, tyrosine, DNA, and Tween 80. Starch, cellulose, and chitin were not degraded. The strains were catalase and nitrate reductase positive, did not produce H2S, and did not grow under anaerobic conditions. F. ovolyticus resembles Flexibacter maritimus, but differs from the latter species in several biochemical and physiological characteristics. DNAs from F. ovolyticus strains had guanine-plus-cytosine contents which ranged from 30.3 to 32.0 mol% (strains EKC001, EKD002T [T = type strain], and VKB004), and DNA-DNA hybridization studies revealed levels of relatedness between F. ovolyticus EKD002T and F. maritimus NCMB 2154T and NCMB 2153 of 42.7 and 30.0%, respectively. Compared with previously described Cytophaga and Flexibacter spp. with low guanine-plus-cytosine contents, F. ovolyticus constitutes a new species. Strain EKD002 (= NCIMB 13127) is the type strain of the new species.

Animals↗

The nematophagous fungus Verticillium chlamydosporium produces a chymoelastase-like protease which hydrolyses host nematode proteins in situ.

The nematophagous fungus Verticillium chlamydosporium secreted several proteases in submerged culture in which soya peptone was the sole carbon and nitrogen source. One protease, VCP1 (M(r) 33,000, pI 10.2), was purified 14-fold from culture filtrates to apparent homogeneity using preparative isoelectric focusing in free solution, and shown to rapidly hydrolyse the chymotrypsin substrate Suc-(Ala)2-Pro-Phe-pNA and elastin. VCP1 had a Km for Suc-(Ala)2-Pro-Phe-pNA of 4.3 x 10(-5) M and a kcat of 5.8 s-1. It was highly sensitive to PMSF and TPCK, but only moderately sensitive to chicken egg-white and soya bean trypsin inhibitors. VCP1 degraded a wide range of polymeric substrates, including Azocoll, hide protein, elastin, casein and albumin, and accounted for most of the non-specific protease activity detected in culture filtrates. The purified enzyme hydrolysed proteins in situ from the outer layer of the egg shell of the host nematode Meloidogyne incognita and exposed its chitin layer. VCP1 was secreted by several isolates of V. chlamydosporium and V. lecanii, pathogens of nematodes and insects respectively, but not plant-pathogenic species of Verticillium. These observations suggest that VCP1 or similar enzyme(s) may play a role in the infection of invertebrates.

Albumins↗

A basic serine protease from Paecilomyces lilacinus with biological activity against Meloidogyne hapla eggs.

Scanning electron micrographs of the nematode-egg-parasitic fungus Paecilomyces lilacinus infecting eggs of the root-knot nematode Meloidogyne spp. suggested the involvement of lytic enzymes. When grown on a liquid mineral salts medium, supplemented with different substrates as the sole N- and C-source, the fungus produced an extracellular protease. Colloidal chitin, vitellin and intact eggs of the root-knot nematode Meloidogyne hapla induced proteolytic activity that was repressed by glucose. The protease was partially purified from the culture filtrate by affinity chromatography. It has a molecular mass of 33.5 kDa, a pH optimum of 10.3, a temperature optimum of 60 degrees C and an isoelectric point above pH 10.2. The enzyme was completely inhibited by PMSF. The amino acid sequence, as derived from the nucleotide sequence of a cDNA clone, had high homology with several subtilisin-like serine proteases. It was shown that the purified enzyme degrades vitellin. The protease quantitatively bound to nematode eggs, and eggs incubated with the purified protease eventually floated. Incubation of the purified protease with nematode eggs significantly influenced their development as demonstrated by time-lapse microscopy. Immature eggs were highly vulnerable to protease treatments, whereas those containing a juvenile were more resistant. In addition, hatched larvae were not visibly affected by the protease. It can be concluded that the serine protease might play a role in penetration of the fungus through the egg-shell of nematodes.

Amino Acid Sequence↗

The Drosophila p38 MAPK pathway is required during oogenesis for egg asymmetric development.

In mammalian cells, the p38 mitogen-activated protein kinase (MAPK) pathway is activated in response to a variety of environmental stresses and inflammatory stimuli. However, the role of p38 MAPK signaling in unchallenged conditions remains largely unknown. We have isolated mutations in a Drosophila p38 MAPKK gene homolog, licorne (lic), and show that during oogenesis, lic is required in the germ line for correct asymmetric development of the egg. In lic mutant egg chambers, oskar mRNA posterior localization is not properly maintained, resulting in anteroposterior patterning defects in the embryo. Furthermore, lic loss-of-function in the germ line leads to reduced EGF receptor activity in dorsal follicle cells and ventralization of the egg shell. Both these defects are associated with a diminution of gurken protein levels in the oocyte. Our phenotypic data argue for a role of lic in a post-transcriptional regulation of the grk gene. Furthermore, they show that in addition to the well-characterized Ras/Raf/ERK MAPK pathway acting in the follicle cells, another related signaling cascade, the p38 MAPK pathway, is required in the germ line for correct axes determination. These results provide the first genetic demonstration of an essential function for a p38 pathway during development.

Amino Acid Sequence↗

Immobilization of Schistosoma mansoni miracidia by activation of the alternate complement pathway at unusually high serum dilution.

Free-swimming Schistosoma mansoni miracidia were immobilized by adding normal mammalian serum to the water. Miracidial immobilizing activity (MIA) was shown to result from activating the alternate pathway of complement (APC). MIA in normal sera was heat-sensitive and antibody independent; it was greatly reduced in factor B-depleted or C6-depleted, but not in C1-depleted, human serum. Addition of purified factor B to B-depleted serum totally restored MIA. Half-maximal MIA in normal human, rabbit, and guinea pig sera was detectable at final dilutions exceeding 1/100, 1/200, and 1/500, respectively, and normal rat serum was particularly potent, with MIA at dilutions exceeding 1/2000. Detection of APC activity at such high dilutions is quite extraordinary and attributed to the hypotonic conditions. We confirmed and extended previous findings that heat-inactivated infection sera also display MIA. Immobilizing activity in irradiated-cercarial vaccine rat serum cofractionated with rat IgG and anti-SWAP antibody activity. Antibody-dependent MIA titres were much lower than for APC-dependent MIA. Based upon light microscope and transmission EM studies, immobilization of miracidia by APC activation was attributed to severe tegumental damage. Miracidia within egg shells were insensitive to MIA.

Animals↗

Interaction of cholera toxin with three life-cycle stages of Schistosoma mansoni: adult worm, egg and cercaria.

We have previously reported that there is an immunological cross-reactivity between Schistosoma mansoni and cholera toxin (CT). In this study, using an immunofluorescence technique with anti-CT antibody, we provide further evidence for this cross-reactivity by demonstrating an antigen, localized in the tegument of S. mansoni adult worms which is cross-reactive with a CT antigen. Anti-CT antibodies also reacted with structures in S. mansoni cercariae and eggs. Additionally, CT itself was found to bind strongly to the gut of the adult worm, gut cells of cercaria and the egg shell. The binding of CT to the parasite was blocked when parasite sections were incubated with CT which had been incubated with the ganglioside GM1. Lipid extraction and isolation of gangliosides demonstrated the presence of GM1 in adult worms. For further analysis of CT-binding structures, the possible interaction of CT with two major schistosome gut antigens, circulating cathodic antigen (CCA) and circulating anodic antigen (CAA), was studied. We found that CT blocked the binding of anti-CCA antibody to the gut of adult worms and that anti-CCA blocked the binding of CT to the worm gut. These findings indicate that CT binds to CCA present in the gut of the parasite and thus has, in addition to GM1, a second binding specificity.

Animals↗

19s and 7s antibody response of Mastomys natalensis in experimental filarial (Litomosoides carinii, Acanthocheilonema viteae, Brugia malayi, B. pahangi) infections.

Specific total antibody (ab), 19s and 7s ab levels in the serum of M. natalensis were investigated after infection with L. carinii, A. viteae, B. malayi and B. pahangi for period of about 500 days p.i., using ELISA (homologous adult antigen) and indirect immunofluorescence tests (IIFT: homologous adult and microfilariae antigen). Total ab levels in L. carinii infected animals rose moderately during prepatency Maximum levels occurred during patency. The response during prepatency was stronger in A. viteae and Brugia spp. infected hosts. Lateron ab levels increased continuously in Brugia infections; in A. viteae infection they decreased with decreasing parasitaemia. 19s abs were stimulated during prepatency and at the beginning of patency, or were found at moderate levels throughout to period of investigation (Brugia infections). 7s abs predominated beginning at the period of late prepatency (IIFT) or at the beginning of patency (ELISA). The time courses of 7s abs corresponded to those of total abs. As obvious by IIFT (adult worm antigen) total and 19s titres were higher against cuticle antigens, egg shell antigens and intrauterine amorphous material than against antigens located in the hypodermis and musculature. 7s abs showed best reactivity with cuticle antigens. Using microfilarial antigens 19s abs reacted predominantly with cuticle antigens whereas 7s abs often showed higher titres against antigens which were localized within the larvae than against cuticle antigens.

Animals↗

Outbreak of disseminated zygomycosis and concomitant pulmonary aspergillosis in breeder layer cockerels.

Disseminated zygomycosis and concomitant pulmonary aspergillosis were diagnosed in breeder layer cockerels. Five- to 9-week-old breeder layer cockerels with a history of an increased mortality rate were presented to a diagnostic laboratory for examination. On necropsy, large, multifocal, firm, tan, nodules were observed in the lungs, air sacs, peritoneum, livers, spleens and kidneys. On histopathology, mixed populations of zygomycetes and Aspergillus hyphae were observed in the granulomas in the lungs, and zygomycete hyphae were observed in the granulomas in the air sacs, peritoneum, livers, spleens and kidneys. No bacteria were isolated from any of the lesions. Aspergillus fumigatus was isolated from the lung lesions only and hyphae that were consistent with those of a Rhizopus spp. were isolated from the lesions in several organs. Pullets, which were kept together with the cockerels from the day they were hatched, were not affected. The absence of infection in the pullets, which were kept together with the cockerels, suggests that the cockerels were either infected during incubation, with the fungi penetrating the egg shell, or that they were infected during hatching before they were mixed with the pullets.

Animals↗